Administration / exposure

Route of administration:

oral: gavage

Vehicle:

- Vehicle(s)/solvent(s) used: corn oil]- Justification for choice of solvent/vehicle: The vehicle was chosen due to its relative non-toxicity for the animals. The administered volume was 10 mL/kg b.w. including test substance.

Details on exposure:

PREPARATION OF DOSING SOLUTIONS:On the day of the experiment, the test item was dissolved in corn oil. All animals received a single standard volume orally.

Examinations

Tissues and cell types examined:

Per animal 2000 polychromatic erythrocytes (PCE) were analysed for micronuclei. To investigate a cytotoxic effect the ratio between polychromatic and normochromatic erythrocytes was determined in the same sample and expressed in polychromatic erythrocytes per 2000 erythrocytes.

Details of tissue and slide preparation:

CRITERIA FOR DOSE SELECTION:A preliminary study on acute toxicity was performed with two animals per sex under identical conditions as in the mutagenicity study concerning: animal strain, vehicle, route, frequency, and volume of administration. The animals were treated orally with the test item and examined for acute toxic symptoms at intervals of approximately 1 h, 2-4 h, 6 h, 24 h, 30 h, and 48 h after administration of the test item.

TREATMENT AND SAMPLING TIMES ( in addition to information in specific fields):At the beginning of the treatment the animals (including the controls) were weighed and the individual volume to be administered was adjusted to the animal’s body weight. The animals received the test item, the vehicle, or the positive control substance once orally. Six males and six females were treated per dose group and sampling time. Five males and five females each were treated for each vehicle and the positive control group. The animals of all dose groups, except the positive control were examined for clinical signs at intervals of around 1 h, 2 - 4 h, 6 h, 24 h, and/or 48 h after administration of the test item and vehicles. Sampling of the bone marrow was done 24 and 48 hours after treatment, respectively.

Evaluation criteria:

The study was considered valid as the following criteria are met:- at least 5 animals per test group can be evaluated.- PCE to erythrocyte ratio should not be less than 20 % of the vehicle control.- the positive control shows a statistically significant and biological relevant increase of micronucleated PCEs compared to the vehicle control.

A test item is classified as mutagenic if it induces either a dose-related increase or a clear increase in the number of micronucleated polychromatic erythrocytes in a single dose group. Statistical methods (nonparametric Mann-Whitney test (8)) are used as an aid in evaluating the results, if necessary. However, the primary point of consideration is the biological relevance of the results.A test item that fails to produce a biological relevant increase in the number of micronucleated polychromatic erythrocytes is considered non-mutagenic in this system.

Statistics:

Statistical methods (nonparametric Mann-Whitney test are used as an aid in evaluating the results, if necessary.

Results and discussion

Test results

Sex:

male/female

Genotoxicity:

negative

Toxicity:

yes

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Additional information on results:

RESULTS OF RANGE-FINDING STUDYTwo animals of each sex treated in the pre-experiments received the test item 2-Butenoic acid, 4-oxo-4-(tridecylamino)-, (Z)-, branched dissolved in corn oil once orally. The volume administered was 10 mL/kg b.w.. A correction factor of 1.09 was applied. On the basis of these data 2000 mg/kg b.w. for the males and 1500 mg/kg b.w. for the females were estimated to be suitable as highest dose level. Gender specific differences in toxicity were observed. In accordance with the test guidelines both sexes of animals were used in the main experiment.

Any other information on results incl. tables

The test item 2-Butenoic acid, 4-oxo-4-(tridecylamino)-, (Z)-, branched was assessed in the micronucleus assay for its potential to induce micronuclei in polychromatic erythrocytes (PCE) in the bone marrow of the mouse.

The test item was dissolved in corn oil, which was also used as vehicle control. The volume administered orally was 10 mL/kg b.w.. 24 h and 48 h after a single administration of the test item the bone marrow cells were collected for micronuclei analysis. A correction factor of 1.09 was applied.

Six males and six females per test group (except the vehicle and positive control groups with 5 males and 5 females each) were evaluated for the occurrence of micronuclei. Per animal 2000 polychromatic erythrocytes (PCEs) were scored for micronuclei.

To investigate a cytotoxic effect due to the treatment with the test item the ratio between polychromatic and normochromatic erythrocytes was determined in the same sample and reported as the number of PCEs per 2000 erythrocytes.

The highest dose levels were estimated by pre-experiments to be suitable.

The animals treated with the test item showed clinical signs such as reduced spontaneous activity, eyelid closure, tumbling, tremor, diarrhoea and/or ruffled fur in the high dose group (2000 and 1500 mg/kg b.w.) and few female mice showed eyelid closure at 375 mg/kg b.w. in the main experiment. One male of the high dose group (animal no. 67) died 24 hours after treatment.After treatment with the test item at 48h preparation interval the number of PCEs per 2000 erythrocytes was not substantially decreased as compared to the mean value of PCEs per 2000 erythrocytes of the vehicle control thus indicating that 2-Butenoic acid, 4-oxo-4-(tridecylamino)-, (Z)-, branched did not induce cytotoxic effects in the bone marrow.

In comparison to the corresponding vehicle controls there was no statistically significant or biologically relevant enhancement in the frequency of the detected micronuclei at any preparation interval and dose level (except mid dose females, discussed below) after administration of the test item. The mean values of micronuclei observed after treatment with all doses of 2-Butenoic acid, 4-oxo-4-(tridecylamino)-, (Z)-, branched were below or near to the value of the vehicle control group.

The micronucleus frequency found in the mid dose group for the females was statistically significantly higher compared to the vehicle control. However, all values observed in the test item treated dose groups at any preparation interval were very well within the laboratory’s historical vehicle control data. Additionally no dose dependence was observed in any gender. Thus, the observed significance was not biologically relevant and considered to be incidental.

40 mg/kg b.w. cyclophosphamide administered orally was used as positive control which showed a statistically significant increase of induced micronucleus frequency.

In conclusion, it can be stated that during the study described and under the experimental conditions reported, 2-Butenoic acid, 4-oxo-4-(tridecylamino)-, (Z)-, branched did not induce micronuclei as determined by the micronucleus test in the bone marrow cells of the mouse.

Applicant's summary and conclusion

Information on Registered Substances comes from registration dossiers which have been assigned a registration number. The assignment of a registration number does however not guarantee that the information in the dossier is correct or that the dossier is compliant with Regulation (EC) No 1907/2006 (the REACH Regulation). This information has not been reviewed or verified by the Agency or any other authority. The content is subject to change without prior notice.Reproduction or further distribution of this information may be subject to copyright protection. Use of the information without obtaining the permission from the owner(s) of the respective information might violate the rights of the owner.

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